Sakamoto K, Hirose H, Onizuka A, Hayashi M, Futamura N, Kawamura Y, Ezaki T
First Department of Surgery, Department of Microbiology, Gifu University School of Medicine, Tsukasa-machi 40, Gifu, 500-8076, Japan.
J Surg Res. 2000 Dec;94(2):99-106. doi: 10.1006/jsre.2000.5937.
Quantification of changes in gastrointestinal morphology and mucus gel has been difficult to study. In the present study, we investigated changes in rat intestine under total parenteral nutrition (TPN) using fluoresceinated lectin staining and image analysis. Wistar rats (n = 34) were divided into two groups: one group received TPN for 2 weeks, and a control group received standard rat chow and water ad libitum for the same period. A 1-cm segment of distal ileum was removed and cut into cross sections. Sections were stained with hematoxylin and eosin, and to stain the mucus, periodic acid-Schiff (PAS), alcian blue (AB), and fluoresceinated lectin, that is, FITC-labeled Ulex europaeus agglutinin I (FITC-UEA-I), were used. Light microscope images were stored in a personal computer and analyzed using image analysis. We measured perimeter length, mucosal thickness, villus area, villus surface area index, mucus stain-positive area, mucosal area ratio, and mucosal surface area ratio. Perimeter length, mucosal thickness, villus area, and villus surface area index in the TPN group were significantly less than those in the control group (P < 0.001 for each parameter). In all mucus stainings, the stain-positive area in the TPN group was significantly less than that in the control group. However, there were no significant differences in mucosal area or mucosal surface area ratios between the two groups. The FITC-UEA-I-positive area was significantly greater than the PAS- or and AB-positive area. There were significant positive correlations between the FITC-UEA-I-positive area and both the PAS-positive and AB-positive areas. TPN for 2 weeks promoted intestinal atrophy and decreased absolute quantity of mucus gel. We successfully introduced the FITC-UEA-I staining method to evaluate changes in mucus gel.
胃肠道形态和黏液凝胶变化的定量研究一直颇具难度。在本研究中，我们采用荧光素标记凝集素染色和图像分析技术，对全肠外营养（TPN）状态下大鼠肠道的变化进行了研究。将34只Wistar大鼠分为两组：一组接受2周的TPN，对照组在同一时期随意给予标准大鼠饲料和水。取1厘米长的回肠末端切段并制成横断面。切片用苏木精和伊红染色，为了对黏液进行染色，使用了过碘酸-希夫（PAS）、阿尔辛蓝（AB）和荧光素标记凝集素，即异硫氰酸荧光素标记的荆豆凝集素I（FITC-UEA-I）。光学显微镜图像存储在个人计算机中，并使用图像分析软件进行分析。我们测量了周长、黏膜厚度、绒毛面积、绒毛表面积指数、黏液染色阳性面积、黏膜面积比和黏膜表面积比。TPN组的周长、黏膜厚度、绒毛面积和绒毛表面积指数均显著低于对照组（各参数P < 0.001）。在所有黏液染色中，TPN组的染色阳性面积均显著低于对照组。然而，两组之间的黏膜面积或黏膜表面积比没有显著差异。FITC-UEA-I阳性面积显著大于PAS或AB阳性面积。FITC-UEA-I阳性面积与PAS阳性面积和AB阳性面积均呈显著正相关。2周的TPN导致肠道萎缩，并使黏液凝胶的绝对量减少。我们成功引入了FITC-UEA-I染色方法来评估黏液凝胶的变化。
